Na+ TRANSPORT INHIBITION BY RESPIRATORY SYNCYTIAL VIRUS

呼吸道合胞病毒对 Na 转运的抑制

• 批准号: 6931548
• 负责人: IAN CHRISTOPHER DAVIS
• 金额: $ 5.75万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-09-30 至 2006-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6931548
• 关键词: Xenopus; gene expression; host organism interaction; ion transport; laboratory mouse; membrane potentials; proteasome; respiratory epithelium; respiratory syncytial virus; sodium ion; tissue /cell culture; ubiquitin; virus infection mechanism; virus protein; voltage /patch clamp

CANDIDATE: I graduated from the University of Bristol in 1992, with Honours in Anatomical Science and Veterinary Science. At UAB I received training in laboratory animal medicine, laboratory animal pathology, and viral pathogenesis in animal models (Ph.D. February 2000; mentor Dr. Patricia Fultz). Subsequently, I elected to join Dr. Matalon's research group, where I might build on my knowledge of viral pathogenesis by cross-mining in electrophysiology, and apply these techniques to study the pathogenesis of human respiratory viral pneumonitides. RESEARCH: Respiratory syncytial virus (RSV) is commonest cause of lower respiratory tract disease in children worldwide. Pathogenesis of RSV-induced bronchiolitis is poorly understood, and effects of RSV infection on ion transport (a seminal function of respiratory epithelial cells) have not been investigated. I hypothesize that RSV infection of respiratory epithelial cells reduces their Na ? transport capacity. Preliminary studies have demonstrated that this hypothesis is correct, both in vitro and in RSV-infeeted BALB/c mice. My aims for years 01- 03 are to: (1) quantify alterations in Na + transport across airway and alveolar epithelia in vivo and ex vivo, after infection of BALB/c mice with RSV; (2) define changes in Na + currents and amiloride-sensitive channel activity after RSV infection of murine epithelial cells in vitro; and (3) correlate alterations in Na + transport induced by RSV in vitro and in vivo with alterations in ENaC expression by marine respiratory epithelia. My plan for years 04-05 is to determine the role of the ubiquitin/proteasome pathway in mediating reduced Na+ transport after RSV infection of respiratory epithelia, and to identify the role of the RSV small hydrophobic (SH) gene product in modulation of Na + transport. I will use a combined electrophysiologic and biochemical approach to investigate effects of RSV on Na + transport at all levels, from the single cell to the whole animal, and to correlate these effects to Na + channel expression and degradation. My project will emphasize cross-training in diverse techniques, including short-circuit measurements across monolayers, radioisotopic ion flux studies, whole cell and single channel patch-clamp, and measurement of alveolar fluid clearance and nasal potential difference in mice. ENVIRONMENT: This SERCA, with Dr. Matalon (lung physiology) as mentor and Dr. Sullender (respiratory syncytial virus) as co-mentor, will provide the training and setting I require for my maturation into an independent scientist focused on comparative pathophysiologic effects of respiratory viruses on normal epithelial cell function.

候选人：我于1992年毕业于布里斯托大学，获得解剖学和兽医学荣誉学位。在UAB，我接受了实验室动物医学、实验室动物病理学和动物模型中病毒发病机制的培训（博士学位）。2000年2月;导师Patricia Fultz博士）。随后，我选择加入Matalon博士的研究小组，在那里我可以通过电生理学的交叉挖掘来建立我对病毒发病机制的知识，并应用这些技术来研究人类呼吸道病毒性肺炎的发病机制。 研究：呼吸道合胞病毒（RSV）是全球儿童下呼吸道疾病的最常见原因。RSV诱导的细支气管炎的发病机制知之甚少，RSV感染对离子转运（呼吸道上皮细胞的精液功能）的影响尚未研究。我假设呼吸道上皮细胞RSV感染减少他们的Na？运输能力。初步研究表明，这一假设是正确的，无论是在体外和RSV感染的BALB/c小鼠。01年的目标- 03是：（1）定量RSV感染BALB/c小鼠后体内和离体Na +跨气道和肺泡上皮转运的变化;（2）确定RSV感染体外鼠上皮细胞后Na +电流和阿米洛利敏感性通道活性的变化;（3）RSV在体内外诱导的Na +转运的改变与海洋呼吸上皮细胞ENaC表达的改变相关。我04-05年的计划是确定呼吸道上皮RSV感染后泛蛋白/蛋白酶体途径在介导Na+转运减少中的作用，并确定RSV小疏水性（SH）基因产物在调节中的作用Na +转运。我会用一个电生理和生化结合的方法 研究RSV对从单个细胞到整个动物的所有水平的Na +转运的影响，并将这些影响与Na +通道表达和降解相关联。我的项目将强调不同技术的交叉训练，包括单层短路测量，放射性同位素离子通量研究，全细胞和单通道膜片钳，以及小鼠肺泡液体清除率和鼻电位差的测量。 环境：这个SERCA，与Matalon博士（肺生理学）作为导师和Sullender博士（呼吸道合胞病毒）作为共同导师，将提供我成长为一名独立科学家所需的培训和环境，专注于呼吸道病毒对正常上皮细胞功能的比较病理生理学影响。